Synthetic bi-directional plant promoter

Patent metadata

Field	Value
Publication number	US-2016130595-A1
Application number	US-201514938643-A
Country	US
Kind code	A1
Filing date	Nov 11, 2015
Priority date	Nov 11, 2014
Publication date	May 12, 2016
Grant date	—

Abstract

This disclosure concerns compositions and methods for promoting transcription of a nucleotide sequence in a plant or plant cell, employing a minimal core promoter element from a Arabidopsis thaliana Ubiquitin-10 gene promoter or Cassava Vein Mosaic Virus promoter, and the full-length nucleotide sequence elements from a Cassava Vein Mosaic Virus promoter. Some embodiments relate to a synthetic CsVMV bi-directional promoter that functions in plants to promote transcription of two operably linked nucleotide sequences.

First claim

What is claimed is:

1 . A synthetic CsVMV bi-directional polynucleotide promoter comprising a plurality of promoter elements from an Arabidopsis thaliana Ubiquitin-10 promoter and a Cassava Vein Mosaic Virus promoter.

2 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the promoter elements comprise a minimal core promoter.

3 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 2 , wherein the minimal core promoter comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:1.

4 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 2 , wherein the minimal core promoter comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:5.

5 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the promoter elements comprise an intron.

6 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 5 , wherein the intron comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:2.

7 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the promoter elements comprise a 5′-UTR.

8 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 7 , wherein the 5′-UTR comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:3.

9 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 7 , wherein the 5′-UTR comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:6.

10 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the promoter elements comprise an upstream promoter element.

11 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 10 , wherein the upstream promoter element comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:4.

12 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 10 , wherein the upstream promoter element comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:7.

13 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the Arabidopsis thaliana Ubiquitin-10 promoter comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:8.

14 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , wherein the Cassava Vein Mosaic Virus promoter comprises a polynucleotide sequence with at least 90% sequence identity to SEQ ID NO:9.

15 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , the synthetic CsVMV bi-directional polynucleotide promoter selected from the group consisting of SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13.

16 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , the synthetic CsVMV bi-directional polynucleotide promoter comprising SEQ ID NO:10.

17 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , the synthetic CsVMV bi-directional polynucleotide promoter comprising SEQ ID NO:11.

18 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , the synthetic CsVMV bi-directional polynucleotide promoter comprising SEQ ID NO:12.

19 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 1 , the synthetic CsVMV bi-directional polynucleotide promoter comprising SEQ ID NO:13.

20 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 15 , comprising a first polynucleotide sequence of interest operably linked to the 3′ end of the synthetic CsVMV bi-directional polynucleotide promoter selected from the group consisting of SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13.

21 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 20 , comprising a second polynucleotide sequence of interest operably linked to the 5′ end of the synthetic CsVMV bi-directional polynucleotide promoter selected from the group consisting of SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13.

22 . The synthetic CsVMV bi-directional polynucleotide promoter as in claims 20 , wherein the polynucleotide sequence of interest comprises a trait.

23 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 22 , wherein the trait is selected from the group consisting of is an insecticidal resistance trait, herbicide tolerance trait, nitrogen use efficiency trait, water use efficiency trait, nutritional quality trait, DNA binding trait, selectable marker trait, and any combination thereof.

24 . A method for producing a transgenic plant cell, the method comprising:
a) transforming a plant cell with a gene expression cassette comprising a synthetic CsVMV bi-directional polynucleotide promoter operably linked to at least one polynucleotide sequence of interest; b) isolating the transformed plant cell comprising the gene expression cassette; and, c) producing a transgenic plant cell comprising the synthetic CsVMV bi-directional polynucleotide promoter operably linked to at least one polynucleotide sequence of interest.

25 . The method of claim 24 , wherein transforming a plant cell is performed with a plant transformation method.

26 . The method of claim 25 , wherein the plant transformation method is selected from the group consisting of an Agrobacterium -mediated transformation method, a biolistics transformation method, a silicon carbide transformation method, a protoplast transformation method, and a liposome transformation method.

27 . The method of claim 24 , wherein the polynucleotide sequence of interest is constitutively expressed throughout the transgenic plant cell.

28 . The method of claim 24 , wherein the polynucleotide sequence of interest is stably integrated into the genome of the transgenic plant cell.

29 . The method of claim 24 , the method further comprising the steps of:
e) regenerating the transgenic plant cell into a transgenic plant; and, f) obtaining the transgenic plant, wherein the transgenic plant comprises the gene expression cassette comprising the synthetic CsVMV bi-directional polynucleotide promoter of claim 1 operably linked to at least one polynucleotide sequence of interest.

30 . The method of claim 24 , wherein the transgenic plant cell is a monocotyledonous transgenic plant cell or a dicotyledonous transgenic plant cell.

31 . The method of claim 30 , wherein the dicotyledonous transgenic plant cell is selected from the group consisting of an Arabidopsis plant cell, a tobacco plant cell, a soybean plant cell, a canola plant cell, and a cotton plant cell.

32 . The method of claim 30 , wherein the monocotyledonous transgenic plant cell is selected from the group consisting of a maize plant cell, a rice plant cell, a Brachypodium plant cell, and a wheat plant cell.

33 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 24 , the synthetic CsVMV bi-directional polynucleotide promoter selected from the group consisting of SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:13.

34 . The synthetic CsVMV bi-directional polynucleotide promoter of claim 33 , comprising a first polynucleotide sequence of interest operably lin

Assignees

• Dow Agrosciences Llc

Inventors

• Kumar Sandeep
• Cicak Toby
• Robinson Heather Leigh
• Pareddy Dayakar Reddy
• Chen Wei

Classifications

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title

• C12N15/8216 Primary

  Methods for controlling, regulating or enhancing expression of transgenes in plant cells · CPC title

• C12N15/8274

  for herbicide resistance · CPC title

• C12N9/1241

  Nucleotidyltransferases (2.7.7) · CPC title

• C12Y207/07047

  Streptomycin 3''-adenylyltransferase (2.7.7.47) · CPC title